Recombinant Rat CXCL5

Recombinant Rat CXCL5 is a valuable tool for research applications focused on inflammation, pain mechanisms, immune cell recruitment, and metabolic disease. Its use enables precise, reproducible studies of CXCL5’s biological functions and signaling pathways in rat models.

Key scientific reasons to use recombinant rat CXCL5 include:

• Modeling Inflammatory Responses: CXCL5 is a potent chemokine that mediates neutrophil and macrophage recruitment during inflammation. Recombinant CXCL5 allows controlled investigation of its role in inflammatory processes, such as those seen in arthritis, UVB-induced skin injury, and other immune-mediated conditions.

• Pain Mechanism Studies: CXCL5 activates CXCR2 on nociceptive sensory neurons, driving pain and hyperalgesia in models of gout and UVB-induced skin injury. Recombinant CXCL5 can be used to induce pain-like behaviors and study underlying molecular mechanisms, including TRPA1 activation and calcium signaling in neurons.

• Cellular Signaling and Migration Assays: CXCL5 triggers calcium responses and promotes migration in macrophages and other immune cells. Recombinant protein is essential for in vitro assays to dissect chemokine-receptor interactions, intracellular signaling, and immune cell dynamics.

• Metabolic Disease Research: CXCL5 regulates adipogenesis and oxidative stress in adipose tissue. Recombinant CXCL5 enables studies on its effects in metabolic pathways, including its role in insulin resistance, ROS regulation, and inflammation in obesity models.

• Standardization and Reproducibility: Using recombinant protein ensures high purity, defined concentration, and batch-to-batch consistency, which are critical for quantitative assays, dose-response studies, and mechanistic experiments.

• Therapeutic Target Validation: Recombinant CXCL5 is used to validate therapeutic strategies targeting the CXCL5/CXCR2 axis, relevant for conditions such as inflammatory pain, metabolic syndrome, and cancer.

Typical applications include:

• In vivo administration to model inflammation or pain in rats.
• In vitro stimulation of immune cells, neurons, or adipocytes.
• Chemotaxis, migration, and calcium flux assays.
• ELISA standards and positive controls in immunoassays.
• Mechanistic studies of CXCL5 signaling pathways.

In summary, recombinant rat CXCL5 is essential for dissecting its biological roles in inflammation, pain, immune cell function, and metabolic regulation, providing a standardized reagent for robust and reproducible research in rat systems.

Yes, recombinant rat CXCL5 can be used as a standard for quantification or calibration in ELISA assays, provided that it is biologically and immunologically equivalent to the native protein detected by your ELISA kit.

Several sources confirm that recombinant rat CXCL5 is suitable for this purpose:

• Novus Biologicals and Bio-Techne both offer recombinant rat CXCL5/ENA-78 proteins that are described as fully biologically active and comparable to standard preparations, making them appropriate for use as ELISA standards (sources , ).
• Thermo Fisher Scientific and Abcam provide ELISA kits for rat CXCL5 that use recombinant standards for calibration, and their protocols recommend preparing dilution series from a stock standard solution (sources , , ).
• The parallelism between recombinant and native proteins is a key validation criterion for ELISA kits, ensuring that the recombinant standard accurately reflects the behavior of the native analyte in the assay (source ).

Best Practices:

• Ensure the recombinant protein is of high purity and has been validated for immunoreactivity with your ELISA antibodies.
• Prepare a dilution series of the recombinant standard in the same matrix as your samples (e.g., assay diluent, serum, or buffer) to generate a standard curve.
• Confirm linearity and parallelism by comparing serial dilutions of your samples to the standard curve.

In summary, recombinant rat CXCL5 is a valid and commonly used standard for ELISA quantification, as long as it is compatible with your assay system and properly validated.

Recombinant Rat CXCL5 has been validated in published research for several key applications, primarily in studies of inflammation, pain, and cancer biology.

Validated Applications in Published Research:

• In vivo functional studies: Recombinant Rat CXCL5 has been used to induce and study inflammatory pain responses in rat skin. Direct injection of CXCL5 into rat skin recapitulated mechanical hypersensitivity and promoted infiltration of neutrophils and macrophages, demonstrating its role as a peripheral mediator of UVB-induced inflammatory pain.

• Cell migration and signaling assays: In vitro, recombinant CXCL5 has been shown to trigger calcium responses and increase migration in cultured macrophages, confirming its chemotactic activity and receptor specificity (primarily via CXCR2).

• Angiogenesis assays: Recombinant CXCL5 has been used to stimulate endothelial cells (e.g., HUVECs) in tube formation assays, demonstrating its ability to promote angiogenesis. In vivo, CXCL5 enhanced angiogenesis in Matrigel plug assays and increased tumor vascularization in xenograft models.

• Cancer progression models: Recombinant CXCL5 has been applied to study its effects on tumor cell proliferation, invasion, and metastasis in various cancer models, including colorectal cancer and other solid tumors. It has been shown to promote tumor growth and angiogenesis, often through the CXCL5/CXCR2 axis.

Additional Context:

• Immunology research: CXCL5 is widely used as a tool to study neutrophil recruitment, inflammatory signaling, and immune cell interactions in rodent models.
• Mechanistic studies: Recombinant CXCL5 is employed to dissect signaling pathways involved in inflammation, tissue remodeling, and tumor microenvironment modulation.

Summary Table:

These applications are supported by peer-reviewed studies and are standard in the characterization of CXCL5’s biological functions in rodent models.

To reconstitute and prepare Recombinant Rat CXCL5 protein for cell culture experiments, follow these best-practice steps:

• Centrifuge the vial briefly before opening to ensure all lyophilized protein is at the bottom.
• Reconstitute the protein in sterile distilled water or an aqueous buffer containing 0.1% BSA (bovine serum albumin) to a final concentration of 0.1–1.0 mg/mL. For example, to make a 0.5 mg/mL stock, add 200 μL of water to 100 μg of protein.
• Gently pipette to dissolve the protein completely, washing down the sides of the vial to recover all material.
• If the protein is difficult to dissolve, allow it to sit on ice for 10–30 minutes with occasional gentle mixing. Avoid vigorous vortexing to prevent denaturation.

Storage after reconstitution:

• Short-term (up to 1 month): Store aliquots at 2–8°C, ideally with a carrier protein such as 0.1% BSA to prevent adsorption and loss of activity.
• Long-term (up to 6 months): Store aliquots at –20°C or –80°C with a carrier protein.
• Avoid repeated freeze-thaw cycles by aliquoting into single-use volumes.

Preparation for cell culture:

• Before adding to cells, dilute the reconstituted stock to the desired working concentration using sterile cell culture medium.
• Typical working concentrations for chemotaxis or signaling assays range from 10–100 ng/mL, but optimal concentrations should be determined empirically for your specific assay.

Additional notes:

• Ensure the endotoxin level is suitable for cell culture (commonly <1 EU/μg).
• If using for sensitive cell types, consider further dilution in medium containing 0.1% BSA or 1–10% serum to stabilize the protein and minimize adsorption to plasticware.

Summary protocol:

1. Centrifuge vial, open carefully.
2. Add sterile water or buffer with 0.1% BSA to achieve 0.1–1.0 mg/mL.
3. Mix gently until fully dissolved.
4. Aliquot and store at 2–8°C (short-term) or –20°C/–80°C (long-term) with carrier protein.
5. Dilute to working concentration in cell culture medium immediately before use.

These steps will ensure the recombinant rat CXCL5 protein is properly prepared and retains biological activity for cell culture experiments.